SUV420H2 suppresses breast cancer cell invasion through down regulation of the SH2 domain-containing focal adhesion protein tensin-3.

The genome-wide loss of histone H4 lysine 20 tri-methylation (H4K20me3) is observed in multiple types of cancer, including breast tumors. Since H4K20me3 is preferentially targeted to repetitive elements in the pericentromeric and telomeric heterochromatin and plays a role in chromatin integrity, the pathological effects of disrupted H4K20me3 in tumors have been attributed to genomic instability. However, in this report, we show that loss of H4K20me3 modulates gene expression profiles, leading to increased cell invasion. Reduced H4K20me3 levels in tumor cells are often accompanied by a decrease in the expression of the H4K20-specific methyltransferase, SUV420H2. Exogenous delivery of SUV420H2 into MDA-MB-231 human breast cancer cells induced selective and specific changes in the expression of cancer-related genes. One of the most downregulated genes in response to SUV420H2 expression was the Src substrate, tensin-3, a focal adhesion protein that contributes to cancer cell migration. Depletion of tensin-3 suppressed breast cancer cell invasiveness. Furthermore, silencing of tensin-3 was associated with enrichment of H4K20me3 immediately upstream of the tensin-3 transcription start site, suggesting that the loss of H4K20me3 in tumor cells induced the expression of cancer-promoting genes. These findings connect the loss of H4K20me3 with tumor progression, through the transcriptional activation of cancer-promoting genes.

Quantification of proteins using (13)C7-labeled and unlabeled iodoacetanilide by nano liquid chromatography/nanoelectrospray ionization and by selected reaction monitoring mass spectrometry.

The combination of cysteine-specific modifiers, iodoacetanilide (IAA) and (13)C7-labeled iodoacetanilide ((13)C7-IAA), has been applied to absolute quantification of proteins. The selected reaction monitoring (SRM) with the use of nano liquid chromatography/nanoelectrospray ionization ion trap mass spectrometry (nano LC/nano-ESI-IT-MS) analysis was applied to precise quantification of three commercial proteins. Good correlation was observed between the theoretical ratios and observed ratios for all these proteins both in a simple buffer solution and in a complex protein environment. Due to efficient tagging, this method does not require separate synthesis of isotope-labeled peptides for the SRM studies. Therefore, this method is expected to be a useful tool for proteomics research.

Cancer-associated upregulation of histone H3 lysine 9 trimethylation promotes cell motility in vitro and drives tumor formation in vivo.

Global histone modification patterns correlate with tumor phenotypes and prognostic factors in multiple tumor types. Recent studies suggest that aberrant histone modifications play an important role in cancer. However, the effects of global epigenetic rearrangements on cell functions remain poorly understood. In this study, we show that the histone H3 lysine 9 (H3K9) methyltransferase SUV39H1 is clearly involved in regulating cell migration in vitro. Overexpression of wild-type SUV39H1, but not enzymatically inactive SUV39H1, activated migration in breast and colorectal cancer cells. Inversely, migration was reduced by knockdown of SUV39H1 or chemical inhibition by chaetocin. In addition, H3K9 trimethylation (H3K9me3) was specifically increased in invasive regions of colorectal cancer tissues. Moreover, the presence of H3K9me3 positively correlated with lymph node metastasis in colorectal cancer patients. Furthermore, overexpression of SUV39H1 drove tumorigenesis in mouse, resulting in a considerable decrease in survival rate. These data indicate that H3K9 trimethylation plays an important role in human colorectal cancer progression, possibly by promoting collective cell invasion.

Analysis of "visible in retrospect" to monitor false-negative findings in radiological reports.

PURPOSE: False-negative findings in radiological reports can lead to serious adverse patient outcomes. We determined the frequency and tendency of false-negative findings in radiological reports by searching for words related to "visible in retrospect". METHODS: In the period of 34 months, we extracted radiological reports containing words related to "visible in retrospect". Of these reports, we extracted false-negative findings that were not described in past reports and were first detected retrospectively. Misinterpretations were excluded. The occurrences of the terms that we identified were analyzed by all examinations, modality, month, and anatomical and lesion classifications were analyzed. RESULTS: Of the 135,251 examinations, 941 reports (0.71%) with 962 findings were detected, with an average of 1.4 findings per business day. By modality, 713 of 81,899 (0.87%) CT examinations, 208 of 36,174 (0.57%) MR, 34 of 9,585 (0.35%) FDG-PET-CT, 2 of 2,258 (0.09%) digital radiography, and 5 of 5,335 (0.09%) other nuclear medicine examinations were found. By anatomical classification, there were 383 (40%) in chest, 353 (37%) in abdomen, 162 (17%) in head, 42 (4.4%) in face and neck, 9 (0.93%) in extremity, and 13 (1.4%) in others. By lesion classification, we identified 665 (69%) for localized lesion, 170 (18%) for vascular lesion, 83 (8.6%) for inflammatory lesion, 14 (1.5%) for traumatic lesion, 12 (1.2%) for organ dysfunction, 11 (1.1%) for degenerative lesion, and 7 (0.7%) for the others. Notable high-frequency specific site diseases by modality were 210 (22%) of localized lesions in lung on CT. CONCLUSION: Our results demonstrated that missed lung localized lesions on CT, which account for about a fifth of false-negative findings, were the most common false-negative finding.

Nuclear envelope-localized EGF family protein amphiregulin activates breast cancer cell migration in an EGF-like domain independent manner.

Amphiregulin (AREG), an EGF family protein, is synthesized as a type I transmembrane precursor (proAREG) and expressed on the cell surface with an extracellular EGF-like domain and an intracellular short cytoplasmic tail. The ectodomain shedding yields a soluble EGF receptor ligand (soluble AREG) which binds to EGF receptor (EGFR) and concomitantly induces migration of unshed proAREG from the plasma membrane to the nuclear envelope (NE). AREG is known to play a potential role in breast cancer and has been intensively investigated as an EGF receptor ligand, while the function of the NE-localized proAREG remains unknown. In this study we used a truncated mutant that mimics NE-localized proAREG without shedding stimuli to discriminate between the functions of NE-localized and plasma membrane-localized proAREG and demonstrate that NE-localized proAREG activates breast cancer cell migration, but suppresses cell growth. Moreover, the present study shows that induction of cell migration by NE-localized proAREG does not require the extracellular growth factor domain or EGF receptor function. Collectively these data demonstrate a novel function mediated by the intracellular domain of proAREG and suggest a significant role for NE-localized proAREG in driving human breast cancer progression.

Dosimetry of Occupational Eye Lens Dose Using a Novel Direct Eye Dosimeter, DOSIRIS, during Interventional Radiology Procedures.

In response to the recommendation by the International Commission on Radiological Protection to lower the equivalent eye dose limit, the Japanese Government in April 2021 lowered the equivalent dose limit for the eye lens for occupational exposure. A considerable number of interventional radiology operators are exposed to levels above the new limit. For this reason, a need exists to more accurately evaluate eye lens dose in interventional radiology operators by using a novel direct eye dosimeter, the DOSIRIS™(IRSN, France), which is capable of measuring a 3-mm dose equivalent under protective glasses. The DOSIRIS is a thermoluminescent dosimeter that exhibits good energy dependence and better directional properties than other dosimeters. Dosimetry using DOSIRIS might be accurate and compatible with the latest regulations.

The SUN2-nesprin-2 LINC complex and KIF20A function in the Golgi dispersal.

The morphology of the Golgi complex is influenced by the cellular context, which strictly correlates with nuclear functions; however, the mechanism underlying this association remains elusive. The inner nuclear membrane SUN proteins, SUN1 and SUN2, have diverse functions together with the outer nuclear membrane nesprin proteins, which comprise the LINC complex. We found that depletion of SUN1 leads to Golgi complex dispersion with maintenance of ministacks and retained function for vesicle transport through the Golgi complex. In addition, SUN2 associates with microtubule plus-end-directed motor KIF20A, possibly via nesprin-2. KIF20A plays a role in the Golgi dispersion in conjunction with the SUN2-nesprin-2 LINC complex in SUN1-depleted cells, suggesting that SUN1 suppresses the function of the SUN2-nesprin-2 LINC complex under a steady-state condition. Further, SUN1-knockout mice, which show impaired cerebellar development and cerebellar ataxia, presented altered Golgi morphology in Purkinje cells. These findings revealed a regulation of the Golgi organization by the LINC complex.

Determination of procalcitonin concentration using the SphereLight 180 clinical auto-analyzer.

BACKGROUND: Procalcitonin (PCT) is a biomarker for the diagnosis of sepsis and bacterial infection diseases. METHODS: A new fully automated SphereLight PCT (SL-PCT) assay system for PCT concentration in human serum or plasma by using SphereLight 180 (SL180, Olympus Corp.) analyzer was developed. The SL-PCT assay is based on chemiluminescent enzyme immunoassay. RESULTS: A linear dose response relationship was observed up to 200 ng/ml PCT concentration. The detection limit of PCT concentration was 0.06 ng/ml. Endogenous substances, anticoagulants, sodium fluoride and drugs did not interfere with assay results. There was a good correlation between the present method and the manual method in serum and plasma samples. CONCLUSIONS: These results indicate that the SL-PCT assay showed good performance in terms of the linearity, detection limit and precision. Use of this PCT measurement may improve the detection of sepsis and infectious disease.

In vitro imaging of human monocytic cellular activity using superparamagnetic iron oxide.

The aim is to evaluate if a difference in activity of monocytic cells can be described on superparamagnetic iron oxide (SPIO)-magnetic resonance imaging (MRI) in vitro when a low concentration of SPIO is administered. Human monocytic cells were stimulated with phorbol 12-myristate 13-acetate (PMA) or left non-stimulated for 24 h and then treated with SPIO with a final concentration of 7.2 microg Fe/ml for 48 h. They were collected, and an MRI phantom was prepared by suspending them in 4% gelatin and scanned using gradient-echo T2*-weighted and spin-echo T2-weighted imaging with two different echo times. The signal intensity and T2*/T2 relaxation times of each layer were also assessed. The number and degree of intracellular filling of cells positively stained with Berlin blue were evaluated microscopically. In addition, the heme oxygenase-1 expression in non-stimulated monocytic cells or monocytic cells stimulated with PMA was evaluated using semiquantitative reverse transcription and polymerase chain reaction. The signal reduction of cells stimulated with PMA was 3.9- to 5.0-fold and 2.7- to 3.1-fold greater than that of non-stimulated cells on the T2*- and T2-weighted images, respectively. These signal reductions were also visually confirmed. Estimated T2*/T2 relaxation times of cells stimulated with PMA and non-stimulated cells were 31.3/137.8 and 122.8/182.6 ms, respectively. Cells positively stained with Berlin blue were observed in 4.5% and 25.3% of non-stimulated cells and those stimulated with PMA, respectively (p<0.05). The intracellular filling in cells stimulated with PMA was significantly larger than that in non-stimulated cells (p<0.05). The expression of the heme oxygenase-1 gene in monocytic cells stimulated with PMA was 2.9-fold greater than that in non-stimulated monocytic cells. SPIO-MRI can visually describe a difference in activity of monocytic cells in vitro when a low concentration of SPIO is administered.

Identification of potential breast cancer markers in nipple discharge by protein profile analysis using two-dimensional nano-liquid chromatography/nanoelectrospray ionization-mass spectrometry.

PURPOSE: This research aimed to establish a diagnostic technique for breast cancer using nipple discharge (ND), with the objective of preventive diagnosis. ND has been proposed as a source of secreted proteomes that reflect early pathological changes in the ductal-lobular epithelial microenvironment, and could thus provide breast-specific cancer biomarkers that could be accessed noninvasively as a new clinical diagnostic technique. EXPERIMENTAL DESIGN: Minute amounts of ND from patients with and without breast cancer (n = 19 and 12, respectively) were collected at the hospital and kept frozen until just before use. They were analyzed using high-pH RP peptide fractionations/low-pH RP 2D nano-LC/ESI-MS/MS. Biomarker candidates were also investigated using Western blot analysis and sandwich ELISA on ND and/or sera. RESULTS: We found distinct tendencies in protein expression and three candidate breast cancer biomarkers (carbonic anhydrase 2, catalase, and peroxiredoxin-2) whose levels differed significantly between ND specimens from patients with and without breast cancer. CONCLUSIONS AND CLINICAL RELEVANCE: These tendencies in protein expression and markers provide new ways to identify breast cancer patients. Therefore, RP/RP 2D LC/MS/MS analyses of ND and the above three markers are supported as a new breast cancer diagnostic technique.

Simultaneous determination of percentage of Lens culinaris agglutinin-reactive alpha-fetoprotein and alpha-fetoprotein concentration using the LiBASys clinical auto-analyzer.

BACKGROUND: Lens culinaris agglutinin (LCA)-reactive alpha-fetoprotein (AFP-L3) percentage of total AFP concentration [(AFP-L3/total AFP)x100] has been used as an effective marker for earlier diagnosis, for assessment of therapeutic effects and for predicting the prognosis of hepatocellular carcinoma (HCC). METHODS: A new clinical automatic analyzer, the "LiBASys", and an assay kit for the simultaneous determination of AFP-L3 percentage and concentration of AFP in human serum were developed. LiBASys performed automatic re-measurement and also met stat samples. Both AFP-L3 percentage and AFP concentration were calculated simultaneously and printed out continuously every 3.4 min per test. RESULTS: A linear dose-response relationship was observed up to 1000 ng/ml AFP concentration. The detection limit and functional sensitivity of LCA-nonreactive AFP (AFP-L1) and AFP-L3 concentrations were 0.4 and 0.8 ng/ml, and 0.4 and 1.0 ng/ml, respectively. Interassay CVs of AFP-L3 percentage and AFP concentration ranged from 2.8% to 13.4% and 2.6% to 4.6%, respectively. Assay results exhibited good correlations with those of commercially available lectin-affinity electrophoresis (r=0.984) for AFP-L3 percentage and with the RIA method (r=0.999) for AFP concentration. CONCLUSIONS: This method simultaneously yields both qualitative and quantitative results for AFP-L3 percentage and AFP concentration.

Morphologic approach to hepatocellular carcinoma development in man: de novo or the so-called 'dysplastic nodule-carcinoma' sequence?

The so-called dysplastic nodule-carcinoma sequence in the liver is generally accepted because hepatocellular carcinoma is not an uncommon finding in precancerous lesions. In order to evaluate the existence and frequency of de novo hepatocarcinogenesis we studied 112 surgically resected early well-differentiated hepatocellular carcinomas showing replacing growth without less differentiated component in themselves. They were divided into two groups: carcinoma in dysplastic area (type A) and carcinoma without dysplastic area (type B) and were analyzed clinicopathologically. We encountered 77 cases of type A (68.8%) and 35 of type B (31.2%). The frequency of type A in cirrhotic group (74.7%) is statistically higher than that of non-cirrhotic group (54.5%) (p=0.0453). Using multivariate analysis, the occurrence of type A was related with higher age, the presence of cirrhosis and hepatitis B surface antigen positive. The tumor size and the presence of fatty change in the tumor tended to relate with type A. We propose two pathways morphologically in early hepatocarcinogenesis, one of which has a close relation to hepatitis B virus and/or cirrhosis.

On the origin of near-infrared extragalactic background light anisotropy.

Extragalactic background light (EBL) anisotropy traces variations in the total production of photons over cosmic history and may contain faint, extended components missed in galaxy point-source surveys. Infrared EBL fluctuations have been attributed to primordial galaxies and black holes at the epoch of reionization (EOR) or, alternately, intrahalo light (IHL) from stars tidally stripped from their parent galaxies at low redshift. We report new EBL anisotropy measurements from a specialized sounding rocket experiment at 1.1 and 1.6 micrometers. The observed fluctuations exceed the amplitude from known galaxy populations, are inconsistent with EOR galaxies and black holes, and are largely explained by IHL emission. The measured fluctuations are associated with an EBL intensity that is comparable to the background from known galaxies measured through number counts and therefore a substantial contribution to the energy contained in photons in the cosmos.

Contrast-enhanced MDCT gastrography for detection of early gastric cancer: Initial assessment of "wall-carving image", a novel volume rendering technique.

OBJECTIVE: We developed a new volume rendering technique, the CT gastrography wall carving image (WC) technique, which provides a clear visualization of localized enhanced tumors in the gastric wall. We evaluated the diagnostic performance of the WC as an adjunct to conventional images in detecting early gastric cancer (EGC). MATERIALS AND METHODS: Thirty-nine patients with 43 EGCs underwent contrast-enhanced MDCT gastrography for preoperative examination. Two observers independently reviewed the images under three different conditions: term 1, Axial CT; term 2, Axial CT, MPR and VE; and term 3, Axial CT, MPR, VE and WC for the detection of EGC. The accuracy of each condition as reviewed by each of the two observers was evaluated by receiver operating characteristic analysis. Interobserver agreement was calculated using weighted-κ statistics. RESULTS: The best diagnostic performance and interobserver agreement were obtained in term 3. The AUCs of the two observers for terms 1, 2, and 3 were 0.63, 0.73, and 0.84, and 0.57, 0.73, and 0.76, respectively. The interobserver agreement improved from fair at term 1 to substantial at term 3. CONCLUSIONS: The addition of WC to conventional MDCT display improved the diagnostic accuracy and interobserver reproducibility for the detection of ECG. WC represents a suitable alternative for the visualization of localized enhanced tumors in the gastric wall.

Description of early gastric cancer with wall-carving technique on multidetector computed tomography.

PURPOSE: We introduce a new volume-rendering method, named the "wall-carving (WC) technique," for luminal organs on contrast-enhanced multidetector row computed tomography (MDCT) using a gas-extension protocol. This study aimed to investigate the enhancement profile of the normal gastric wall, particularly focused on the relation between depth and gray value in the normal gastric wall, as a background to distinguish early gastric cancer (EGC) on MDCT. MATERIALS AND METHODS: We retrospectively evaluated 32 data sets of abdominal CT with EGC. All data sets were obtained with a 64-detector-row CT after administration of an effervescent agent, anticholinergic drug, and intravenous contrast material. The enhancement profile was analyzed in the normal gastric body and antrum (n = 32) by the depth from the air-mucosa interface. We also examined the visual accessibility of the WC image. Institutional review board approval was obtained for this study. RESULTS: The profile curve showed that the thickness of the normal gastric wall was 3.75-6.25 mm (mean 4.8 mm), and the maximum enhancement was observed at the layer of 1.875 mm in each case. The enhancement pattern of the EGC was visually accessible, especially at a depth of 1.875-3.75 mm. CONCLUSION: The enhancement profile showed a peak at the depth within the inner half-thickness of the estimated normal gastric wall. The WC image seems to be a good alternative for detecting tumor enhancement and its relation with the surrounding vascular structures in the gastric wall.

Gastric mucosal changes caused by Lugol's iodine solution spray: endoscopic features of 64 cases on screening esophagogastroduodenoscopy.

AIM: To clarify the endoscopic mucosal change of the stomach caused by Lugol's iodine solution spray on screening esophagogastroduodenoscopy (EGD). METHODS: Sixty-four consecutive patients who underwent EGD for esophageal squamous cell carcinoma screening were included in this study. The records for these patients included gastric mucosa findings before and after Lugol's iodine solution was sprayed. The endoscopic findings of the greater curvature of the gastric body were retrospectively analyzed based on the following findings: fold thickening, exudates, ulcers, and hemorrhage. RESULTS: Mucosal changes occurred after Lugol's solution spray totally in 51 patients (80%). Fold thickening was observed in all 51 patients (80%), and a reticular pattern of white lines was found on the surface of the thickened gastric folds found in 28 of the patients (44%). Exudates were observed in 6 patients (9%). CONCLUSION: The gastric mucosa could be affected by Lugol's iodine; the most frequent endoscopic finding of this effect is gastric fold thickening, which should not be misdiagnosed as a severe gastric disease.

Carboplatin and etoposide combined with radiotherapy for limited-stage small-cell esophageal carcinoma: three cases and review of the literature.

PURPOSE: Small-cell esophageal carcinoma (SCEC) is a rare disease for which standard therapy has not yet been established. We report the results of three cases of limited-stage SCEC treated with combination therapy using carboplatin (CBDCA) and etoposide (VP-16) and radiotherapy. MATERIALS AND METHODS: The clinical stage according to the Japanese Classification of Esophageal Cancer 7th ed. was stage III in 2 cases and stage IVa in 1. These patients with limited-stage SCEC were treated at our institution with four cycles of CBDCA and VP-16, either concurrent with radiotherapy for the second two cycles (n = 2) or followed by radiotherapy after the last cycle (n = 1). RESULTS: A complete response (CR) was obtained for all three patients, resulting in a response rate of 100%. Two patients are alive at 16.4 and 22.5 months after initial treatment. One patient died with myeloid leukemia at 43.5 months after initial treatment. None of the patients had loco-regional recurrence. Brain metastasis was detected in one patient at 7 months after initial therapy and was treated with stereotactic radiotherapy combined with whole brain irradiation. CONCLUSION: CBDCA and VP-16 in combination with radiotherapy should be considered an important treatment option for SCEC.

Effective palliative radiotherapy in primary malignant melanoma of the esophagus: a case report.

INTRODUCTION: Primary malignant melanoma of the esophagus is a rare but highly aggressive tumor with poor prognosis. Surgical resection is the treatment of choice. However, some cases may be diagnosed with advanced inoperable disease. Palliative radiotherapy may be used to relieve symptoms caused by the esophageal tumor. CASE PRESENTATION: We report on a case of advanced inoperable primary malignant melanoma of the esophagus treated with palliative radiotherapy. The patient's dysphagia resolved with radiotherapy. CONCLUSION: Malignant melanoma of the esophagus is rare. Patients with advanced inoperable malignant melanomas of the esophagus benefit from radiation therapy. Radiation therapy is effective for palliation.